Andrew Doherty (A.Doherty at Bris.ac.uk) wrote:
} HI everyone - a simple question. When cells are fixed in 3%
} paraformaldehyde, is the integrity of the membrane maintained? Could I
} detect surface expressed receptors with antibodies without seeing the
} intracellular pools?
Um, practically, no the integrity is not maintained. OK, so we always
permeabilize as well as PFA to see intracellular antigens, but that's more
to be absolutely sure they're full of holes. I've looked at actin
staining in PFA-alone fixed cells and seen it. (actin, that is :-))
I suppose lower concs of PFA might maintain memb. integrity, and I
couldn't possibly comment on glutaraldehyde.
R
--
Richard P. Grant MA DPhil University of Oxford
Nuffield Department Obstetrics and Gynaecology FFPGP
http://users.ox.ac.uk/~lady0266http://www.molbiol.ox.ac.uk/~rgrant
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